Departament de Bioquímica i Biotecnologia, Universitat Rovira i Virgili, Marcel·lí Domingo, s/n 43007, Tarragona, Spain.
Br J Nutr. 2012 Nov 14;108(9):1562-73. doi: 10.1017/S0007114511007057. Epub 2012 Jan 6.
Macrophages play an important role in immunogenic challenges by producing reactive oxygen species, NO and proinflammatory cytokines that can aggravate and propagate local inflammation. Multiple mechanisms regulate these inflammatory processes. NF-κB and activator protein 1 pathways are crucial in the expression of proinflammatory genes, such as TNF-α, IL-1 (α or β) and -6. Some polyphenols, which are present in beverages, vegetables and fruits, and PUFA, which are present in marine oils and fish food, possess anti-inflammatory effects in vivo and in vitro. Our aim in the present study was to assess whether polyphenols and PUFA have synergistic anti-inflammatory effects in murine macrophages in vitro. Inflammation in RAW 264.7 macrophages was induced by lipopolysaccharide at 100 ng/ml. The treatments with molecules were performed by co-incubation for 19 h. A NO production assay by Griess reaction, a phosphoprotein assay by Pathscan ELISA kit and gene expression analysis using the TaqMan® Low-density Array for ninety-one genes related to inflammation, oxidative stress and metabolism were performed to assess the synergistic anti-inflammatory effects of polyphenols, epigallocatechin gallate and resveratrol (Res; 2·5 μg/ml), and the PUFA, DHA and EPA (30 μm). Adding Res+EPA had an enhanced anti-inflammatory effect, in comparison with EPA and Res alone, leading to decreased NO levels; modulating the phospho-stress activated protein kinase/Jun N-terminal kinase (P-SAPK/JNK) level; down-regulating proinflammatory genes, such as IL, chemokines, transcription factors; and up-regulating several antioxidant genes. Therefore, this combination has a stronger anti-inflammatory effect than either of these molecules separately in RAW macrophages.
巨噬细胞通过产生活性氧物质、NO 和促炎细胞因子在免疫原性挑战中发挥重要作用，这些物质可以加重和传播局部炎症。多种机制调节这些炎症过程。NF-κB 和激活蛋白 1 途径在促炎基因如 TNF-α、IL-1（α 或β）和-6 的表达中至关重要。存在于饮料、蔬菜和水果中的某些多酚以及存在于海洋油和鱼类食物中的多不饱和脂肪酸 (PUFA) 在体内和体外具有抗炎作用。本研究的目的是评估多酚和 PUFA 在体外对小鼠巨噬细胞是否具有协同抗炎作用。通过 100ng/ml 的脂多糖诱导 RAW 264.7 巨噬细胞炎症。通过共孵育 19 小时进行分子处理。通过格里斯反应进行 NO 产生测定、Pathscan ELISA 试剂盒进行磷酸蛋白测定以及使用 TaqMan®低密度阵列进行与炎症、氧化应激和代谢相关的 91 个基因的表达分析，以评估多酚、表没食子儿茶素没食子酸酯和白藜芦醇 (Res；2.5μg/ml) 和 PUFA、DHA 和 EPA(30μm) 的协同抗炎作用。与 EPA 和 Res 单独添加相比，添加 Res+EPA 具有增强的抗炎作用，导致 NO 水平降低；调节磷酸化应激激活蛋白激酶/Jun N-末端激酶 (P-SAPK/JNK) 水平；下调促炎基因，如 IL、趋化因子、转录因子；并上调几种抗氧化基因。因此，这种组合在 RAW 巨噬细胞中比这些分子中的任何一种单独使用都具有更强的抗炎作用。
